Genipin crosslinked bioactive collagen/chitosan/hyaluronic acid injectable hydrogels structurally amended via...

Collagen, chitosan and hyaluronic acid based multicomponent injectable and in situ gellating biomimetic hybrid materials for bone tissue engineering applications were prepared in one-step procedure. The bioactive phase in the form of surface-modified silica particles was introduced to the solutions of biopolymers and simultaneously crosslinked with genipin both the biopolymer matrix and dispersed particles at 37 °C. The novel approach presented here involved the use of silica particles which surfaces were priory functionalized with amino groups. That modification makes possible the covalent attachment of silica particles to the polymeric hydrogel network on crosslinking with genipin. That methodology is especially important as it makes possible to obtain the hybrid materials (biopolymer-silica particles) in which the problems related to the potential phase separation of mineral particles, hindering their in vivo application can be eliminated. The hybrids of various compositions were obtained and their physicochemical and biological properties were determined. The in vitro experiments performed under simulated body fluid conditions revealed that the amino-functionalized silica particles covalently attached to the biopolymeric network are still bioactive. Finally, the in vitro cell culture studies shown that the materials developed are biocompatible as they supported MG-63 cells adhesion, proliferation as well as Alkaline phosphatase (ALP) expression.